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Research Communications

Effects of dietary supplementation


with sea buckthorn (Hippophae
rhamnoides) seed and pulp oils on
atopic dermatitis
Baoru Yang,* Kirsti O. Kalimo, Leena M. Mattila, Sinikka E. Kallio,
Jouko K. Katajisto, Olli J. Peltola,# and Heikki P. Kallio*
*Department of Biochemistry and Food Chemistry, University of Turku, Turku, Finland;

Department of Dermatology, University of Turku, Turku, Finland; The Finnish Student Health
Service, Turku, Finland; Turku Polytechnic, Social and Health Care, Turku, Finland;

Department of Statistics, University of Turku, Turku, Finland; and #Central Laboratory,
University Central Hospital of Turku, Turku, Finland

A placebo-controlled, double-blind study was conducted to investigate the effects of seed and pulp oils of sea
buckthorn (Hipphophae rhamnoides) on atopic dermatitis. Linoleic (34%), -linolenic (25%), and oleic (19%)
acids were the major fatty acids in the seed oil, whereas palmitic (33%), oleic (26%), and palmitoleic (25%) acids
were the major fatty acids in the pulp oil. The study group included 49 atopic dermatitis patients who took 5 g
(10 capsules) of seed oil, pulp oil, or paraffin oil daily for 4 months. During follow-up dermatitis improved
significantly in the pulp oil (P 0.01) and paraffin oil (P 0.001) groups, but improvement in the seed oil group
was not significant (P 0.11). Supplementation of seed oil increased the proportion of -linolenic acid in plasma
neutral lipids (P 0.01), and increases of linoleic, -linolenic, and eicosapentaenoic acids in plasma
phospholipids were close to significant (0.05 P 0.1). Pulp oil treatment increased the proportion of
palmitoleic acid (P 0.05) and lowered the percentage of pentadecanoic acid (P 0.01) in both plasma
phospholipids and neutral lipids. In the seed oil group, after 1 month of supplementation, positive correlations
were found between symptom improvement and the increase in proportions of -linolenic acid in plasma
phospholipids (Rs 0.84; P 0.001) and neutral lipids (Rs 0.68; P 0.02). No changes in the levels of
triacylglycerols, serum total, or specific immunoglobulin E were detected. In the pulp oil group, a significant
(P 0.05) increase in the level of high density lipoprotein cholesterol, from 1.38 to 1.53 mmol/L was observed
(J. Nutr. Biochem. 10:622 630, 1999) Elsevier Science Inc. 1999. All rights reserved.
.

Keywords: atopy; Hippophae rhamnoides; plasma fatty acids; plasma lipids; sea buckthorn oils

Introduction typical distribution of lesions. Although genetic factors play


an important role, disturbed epidermal barrier function,
The characteristic signs of atopic dermatitis (AD) include aberrant immune response, and increased production of
dry, scaly, itchy skin with eczematous inflammation and immunoglobulin E (IgE) are actively involved in the pro-
cess of disease development.1 Traditionally, AD therapy
includes the liberal use of topical emollients, avoidance of
The authors acknowledge the financial support of the Center for Interna- skin irritating factors, topical glucocorticoids, systemically
tional Mobility (CIMO), Helsinki, Finland.
Address correspondence to Dr. Heikki P. Kallio, Department of Biochem- administered antihistamines, and antimicrobials. Because
istry and Food Chemistry, University of Turku, FIN-20014 Turku, Finland. evidence of the side effects of steroids and certain antihis-
Received February 23, 1999; accepted July 1, 1999. tamines is steadily accumulating, numerous efforts have

J. Nutr. Biochem. 10:622 630, 1999


Elsevier Science Inc. 1999. All rights reserved. 0955-2863/99/$see front matter
655 Avenue of the Americas, New York, NY 10010 PII S0955-2863(99)00049-2
Sea buckthorn oils and atopic dermatitis: Yang et al.
Table 1 Major fatty acids in sea buckthorn seed and pulp oil (weight percentages)

Fatty acids (%)


Oils 16:0 16:1(n-7) 18:0 18:1(n-9) 18:1(n-7) 18:2(n-6) 18:3(n-3)

Seed oil 11.3 4.4 2.6 18.9 3.2 34.1 24.9


Pulp oil 33.4 24.9 1.0 26.2 7.3 5.1 1.6

gone into the search for treatment alternatives including on high density lipoprotein (HDL) has been reported in the
dietary management of the disease.2 6 experimental model of hyperlipemia in chickens.35
Polyunsaturated fatty acids (PUFA) are important parts Because of the fatty acid compositions and the reported
of the sphingolipids that constitute the water barrier of the effects of the two oils on skin and the immune system, we
epidermis.7 As essential components of cell membranes, designed a double-blind, placebo-controlled clinical trial to
PUFA affect the functions of receptors, enzymes, ion test the effects of sea buckthorn seed oil and pulp oil on AD.
channels, and other messenger systems.8 Essential fatty The severity and change of AD symptoms, fatty acid
acids and their metabolites also participate in the regulation compositions of plasma lipid fractions, levels of cholesterol,
of immune reactions and the inflammatory process.8 10 triacylglycerols, serum total, and specific IgE were exam-
Since the observation of abnormal metabolism of essen- ined simultaneously.
tial fatty acids in AD patients, many investigators have
administered oils of different origins systemically to pa-
tients for treatment. Clinical improvement has been noted Materials and methods
by dietary supplementation of both plant seed oils rich in The study was approved by the Ethical Committee of Turku
n-6 PUFA and fish oil rich in n-3 PUFA.2 6,1113 However, University Central Hospital. The purpose of the study was ex-
contradictory results also have been reported.14,15 More plained to all patients and written consent were given by all subjects.
clinical investigations are needed to verify the effects of
PUFA on AD. Oils
Although the nutritional effects of PUFA have been The seeds and soft parts (berry flesh and peel) were separated from
proved, the role of monounsaturated fatty acids, especially the dried press residue of sea buckthorn juice processing. Seed oil
that of palmitoleic acid (16:1n-7) remains unknown. In- was extracted from seeds and pulp oil from the soft parts by aseptic
creased levels of palmitoleic acid have been found in the supercritical carbon dioxide process.36 The fatty acid compositions
tissues under special conditions such as deficiency of of the two oils analyzed as methyl esters with gas chromatography
essential fatty acids and fatty liver.16 18 The mechanism are shown in Table 1. The oils were encapsulated in soft gelatine
behind this phenomenon is not clear. In a clinical trial capsules each containing 500 mg oil and camouflaged with red and
conducted with 14 subjects, a macadamia nut enriched diet black iron paste (E172). Paraffin oil as placebo was encapsulated
containing palmitoleic acid significantly decreased the analogously. D--tocopherol 0.5 mg was added to each capsule.
The capsules were sealed in plastic jars, coded randomly, and kept
plasma total cholesterol, low density lipoprotein (LDL) at 4C until used.
cholesterol, and triacylglycerol concentrations in plasma.19
Berries of sea buckthorn (Hippophae rhamnoides L.) have
Study design and subjects
been used in Tibetan, Mongolian, and Chinese traditional
medicines for the treatment of different diseases for more The study was designed as a placebo-controlled, parallel, random-
than 1,000 years.20 Oil obtained from the seeds of sea ized, double-blind experiment and was carried out during the
buckthorn berries contains linoleic acid (18:2n-6; 34 40%) period of 1996 to 1998 at the Department of Dermatology,
and -linolenic acid (18:3n-3; 2336%) in abundance.2123 University of Turku and The Finnish Student Health Service,
Turku, Finland. All 78 participating patients had a history of AD
The flesh/peel press residue of the berries after juice from childhood with persistent symptoms during the last 6 months.
processing is rich in pulp oil, which has an exceptionally The participants were randomly divided into three groups receiv-
high palmitoleic acid content (24 39%).21,24,25 ing sea buckthorn seed oil, sea buckthorn pulp oil, or paraffin oil.
Topical applications of the two oils on burned, scaled, Ten oil capsules per day were prescribed, and the whole treatment
wounded, and radioactively damaged skins of both humans period lasted 4 months. Patients were asked to follow their normal
and experimental animals have shown healing and anti- diet throughout the trial and were allowed to use emollients,
inflammatory effects.26 29 Sea buckthorn seed oil is known hydrocortisone cream, and peroral antihistamine as needed. The
to increase the specific and nonspecific immune functions severity of AD was evaluated based on the extent and severity of
of experimental animals.30 32 Dietary supplementation with AD symptoms. For evaluation, the SCORAD system37 was
seed oil also has been shown to have a protective effect on adopted in adult measures with a range in score from 0 (no
symptoms) to 3 (extensive severe symptoms). Pruritus and sleep
cell membranes against lipid peroxidation in animal exper- loss were evaluated separately. The maximum total SCORAD
imental models.33,34 value was 103. Patients were clinically examined at the beginning
Sea buckthorn seed oil has been administered to both of the trial, after 1 month, and at 4 months, which was the end of
humans and experimental animals with hyperlipemia, indi- the trial. During each visit, consumption of the capsules was
cating lowering of the high levels of total cholesterol and controlled using oral communication, and any unconsumed cap-
triacylglycerol in plasma.35 An increasing effect of the oil sules were returned by the patients at the end of the trial. During

J. Nutr. Biochem., 1999, vol. 10, November 623


Research Communications
Table 2 Symptom SCORAD values of atopic dermatitis in the three groups at different time points during the administration period

A B C
Groups n mean SD mean SD mean SD

Sea buckthorn seed oil 11 28.0 15.5 22.7 17.1 22.0 17.1
Sea buckthorn pulp oil 16 37.2 17.7 29.2* 20.8 26.8* 19.8
Placebo 18 40.3 16.6 28.4* 14.2 20.6* 16.7

*Significant (P 0.01) compared with point A.


Aat the starting point of the trial. Bafter 1 month of treatment. Cafter 4 months of treatment.

all visits, plasma and serum samples were taken from the patients effect, time effect, and group time interaction effect. Observed
for analysis of the fatty acids of plasma phospholipids and neutral significance levels of less than 0.05 were considered statistically
lipids and levels of cholesterol, triacylglycerols, serum total, and significant; levels of less than 0.1 were considered nearly signifi-
specific IgE. cant. Exact P-values are reported in the text. Correlations between
changes in SCORAD values and changes in proportions of fatty
Analysis of fatty acid compositions of plasma acids in plasma lipids were calculated with the Spearman correla-
tion coefficient. To perform an overall test for the correlations
phospholipids and neutral lipids between symptom improvement and fatty acid changes in plasma
Lipids were extracted from 2 g of plasma with 12 mL chloroform- lipids, the correlation coefficients were tested using the Wilcoxon
methanol (2:1, v/v) using a modified Folch procedure.38,39 The Signed Rank Test by SAS 6.12 Univariate Procedure.
lipids were fractionated on silica Sep-Pak columns by eluting with
10 mL chloroform (neutral lipids) and 20 mL methanol (phospho-
lipids). The glycerophospholipids and neutral lipids were transes- Results
terified by sodium methoxide catalysis.40 The fatty acid methyl
esters (FAME) were analyzed with a Perkin Elmer AutoSystem Patients and power of the study
Gas Chromatograph equipped with programmed split/splitless
injector and flame ionization detector and controlled with the
During follow-up, 29 patients were excluded due to irreg-
Turbochrom Navigator 4 (Perkin Elmer, San Jose, CA USA). ular (or incomplete) use of the capsules, unscheduled
Silica capillary gas chromatography (GC) column NB-351 (L plasma sampling, or clinical examination. The remaining 49
25 m, inner diameter 0.32 mm, df 0.2 m) was used for GC patients, (16 male and 33 female) were included in the data
analysis (HNU-Nordion Ltd, Helsinki, Finland). Flow rate of the analyses. Adequate sample sizes were calculated using
carrier gas helium was 1.7 mL/min, and the split valve with a split SOLO Power Analysis software (BMDP Statistical Soft-
ratio of 1:40 was opened after 1 minute. The temperature program ware Inc., Los Angeles, CA USA, 1991). The minimum
was 120C held for 2 minutes, increased at a rate of 3C/min to sample size in repeated measure ANOVA tests was 5
230C, and held for 20 minutes. The injector temperature was subjects (in levels of antigen-specific IgEs in the seed oil
programmed from 170C to 250C at a rate of 200C/min. The group), and the power of detecting interaction effect was
detector temperature was 270C. FAME were identified by com-
parison with a standard mixture of known composition (68D,
0.71. There were usually at least 12 subjects in each group,
NuChek Prep, Elysian, MN USA) and the fatty acid composition and the power of testing interaction effect was over 0.9.
was expressed as weight percentage of the total fatty acids. The
relative contents of fatty acids of the standard mixture agreed with Effects on symptom severity
the values stated by the manufacturer, with deviations of less than 5%.
The mean SCORAD values are shown in Table 2. Com-
Determination of plasma lipid and IgE levels pared with baseline values, the mean SCORAD values
representing symptom severity were significantly lower
Levels of total cholesterol and triacylglycerols in plasma were after 1-month and 4-month administration in the pulp oil
determined with the reagents CHOD-PAP and GPO-PAP (Boehr- and placebo groups. In the seed oil group, the decrease in
inger Mannheim, Mannheim, Germany), respectively, and anal-
the value was not significant (P 0.11).
ysed with Hitachi 704 and 717 automatic analyzers (Hitachi Ltd,
Tokyo, Japan). HDL cholesterol was determined with Hitachi 704
and 717 automatic analyzers after PEG-6000 precipitation accord- Effects on fatty acid composition of plasma
ing to Izzo et al.41 LDL cholesterol levels in plasma were phospholipids
calculated with the formula of Friedewald et al.42 The radioimmu-
noassay method43 was used for the determination of serum total Twenty-one fatty acids were identified in the plasma phos-
IgE level. The IgE antibodies specific to Saccharomyces cerevi- pholipids, and the results are shown in Table 3. The
siae, Candida albicans, and Pityrosporon orbiculare were deter- proportion of -linolenic acid in the seed oil group in-
mined with CAP-RAST methods.44 creased after 1 month of administration (P 0.08), and
remained stable for the next 3 months. The proportions of
Statistical analysis linoleic acid and eicosapentaenoic acid (20:5n-3) were
Data analysis was carried out by statistical program packages increased by seed oil treatment almost significantly
Statistic/W version 4.5 (Stat Soft Inc., Tulsa, OK USA) and SPSS (0.05 P 0.1). Supplementation for 1 month with pulp
7.5 (SPSS Inc., Chicago, IL USA). Repeated measures analysis of oil clearly increased the proportion of palmitoleic acid of
variance (ANOVA) were used to calculate significance of group phospholipid fatty acids (P 0.05). The percentage of

624 J. Nutr. Biochem., 1999, vol. 10, November


Table 3 Weight percentage of the main fatty acids in the plasma phospholipids of patients in the three groups at different time points during administration

Sea buckthorn seed oil Sea buckthorn pulp oil Placebo


A B C A B C A B C
Fatty acids n Mean SD Mean SD Mean SD n Mean SD Mean SD Mean SD n Mean SD Mean SD Mean SD

14:0 12 0.37 0.43 0.26 0.08 0.76 1.69 16 0.30 0.11 0.29 0.10 0.31 0.12 21 0.26 0.06 0.28 0.11 0.28 0.08
15:0 11 0.17 0.00 0.16 0.03 0.16 0.04 16 0.18 0.03 0.15 0.00 0.16 0.03 21 0.16 0.05 0.18 0.04 0.18 0.05
16:0 12 26.58 1.79 26.00 2.31 25.53 2.43 16 26.38 2.51 25.38 2.52 26.80 1.73 21 25.24 2.26 25.79 1.06 25.73 1.62
16:1 (n-7) 12 1.08 1.75 0.70 0.33 0.60 0.19 16 0.67 0.04 0.86 0.25 0.81* 0.22 21 0.53 0.19 0.54 0.17 0.57 0.13
18:0 12 12.13 1.43 12.51 1.33 12.45 1.16 16 11.75 1.49 10.91 1.47 11.74 1.22 21 12.46 1.32 12.39 1.31 12.46 1.33
18:1 (n-9) 12 10.17 0.90 10.90 1.28 10.54 1.44 16 11.25 1.25 11.74 2.04 11.79 2.12 21 10.30 1.30 10.60 1.25 10.78 1.31
18:1 (n-7) 12 1.81 0.27 1.75 0.25 1.76 0.23 16 1.93 0.21 1.92 0.25 2.00 0.30 21 1.84 0.22 1.80 0.16 1.80 0.18
18:2 (n-6) 12 22.44 2.55 22.55 2.81 23.89* 2.57 16 21.99 3.15 21.10 2.99 23.21 3.01 21 22.77 2.88 22.61 2.27 21.72 5.42
18:3 (n-6) 6 0.09 0.09 0.10 0.09 0.11 0.11 8 0.06 0.03 0.05 0.03 0.08 0.03 12 0.05 0.01 0.06 0.02 0.06 0.03
18:3 (n-3) 12 0.29 0.07 0.37* 0.14 0.37* 0.14 16 0.31 0.13 0.31 0.14 0.34 0.13 21 0.32 0.15 0.32 0.12 0.30 0.08
18:4 (n-3) 12 0.17 0.04 0.17 0.05 0.15 0.04 16 0.20 0.06 0.20 0.06 0.18 0.05 21 0.16 0.05 0.17 0.04 0.16 0.05
20:0 10 0.04 0.01 0.05 0.01 0.05 0.01 11 0.04 0.01 0.04 0.01 0.05 0.01 16 0.05 0.01 0.04 0.01 0.05 0.01
20:1 (n-9) 12 0.16 0.04 0.15 0.04 0.16 0.04 12 0.16 0.03 0.15 0.03 0.15 0.05 21 0.17 0.06 0.15 0.03 0.16 0.03
20:2 (n-6) 12 0.37 0.08 0.34 0.05 0.36 0.07 16 0.38 0.09 0.38 0.01 0.38 0.12 21 0.35 0.08 0.35 0.10 0.38 0.11
20:3 (n-6) 12 3.65 0.91 3.45 0.67 3.29 0.77 16 3.12 0.76 3.03 0.63 3.10 0.56 21 2.99 0.66 3.00 0.68 3.01 0.81
20:4 (n-6) 12 9.76 2.50 9.45 1.93 9.52 2.16 16 9.01 2.03 9.01 1.79 8.72 1.49 21 9.74 1.43 9.85 1.58 9.78 1.77
20:5 (n-3) 12 1.19 0.40 1.41* 0.5 1.30 0.59 16 1.18 0.62 1.16 0.71 1.21 0.56 21 1.61 1.60 1.38 0.40 1.48 0.88
22:4 (n-6) 12 0.28 0.07 0.27 0.07 0.26 0.06 16 0.27 0.08 0.30 0.08 0.27 0.07 20 0.27 0.06 0.26 0.05 0.26 0.05
22:5 (n-6) 12 0.19 0.06 0.18 0.06 0.17 0.06 16 0.19 0.12 0.19 0.09 0.18 0.07 21 0.17 0.07 0.18 0.09 0.19 0.10
22:5 (n-3) 12 0.92 0.23 0.93 0.25 0.92 0.26 16 0.89 0.27 0.85 0.08 0.88 0.26 21 0.88 0.20 0.95 0.28 0.88 0.21
22:6 (n-3) 12 4.26 1.71 4.13 1.97 3.92 2.81 16 4.29 1.35 3.91 1.50 3.88 1.56 21 4.66 1.56 4.81 1.62 4.70 1.61

*Almost significant (0.05 P 0.1) compared with point A.



Significant (P 0.05) compared with point A.

Significant (P 0.01) compared with point A.
Aat the starting point of the trial. Bafter 1 month of treatment. Cafter 4 months of treatment.

J. Nutr. Biochem., 1999, vol. 10, November


625
Sea buckthorn oils and atopic dermatitis: Yang et al.
626
Table 4 Weight percentage of main fatty acids in the plasma neutral lipids of the patients in the three groups at different time points during administration
Research Communications

Sea buckthorn seed oil Sea buckthorn pulp oil Placebo


A B C A B C A B C
Fatty acids n Mean SD Mean SD Mean SD n Mean SD Mean SD Mean SD n Mean SD Mean SD Mean SD

14:0 12 1.77 1.21 1.57 0.91 1.72 0.66 16 1.62 0.81 1.64 0.68 1.47 0.61 21 1.39 0.50 1.40 0.46 1.49 0.63
14:1 (n-5) 12 0.15 0.16 0.12 0.09 0.12 0.06 15 0.16 0.11 0.15 0.09 0.13 0.08 18 0.10 0.06 0.11 0.05 0.12 0.06
15:0 12 0.24 0.08 0.23 0.08 0.23 0.06 16 0.24 0.05 0.21* 0.04 0.20* 0.04 21 0.23 0.05 0.22 0.06 0.23 0.06
16:0 12 19.11 5.39 18.17 4.35 19.05 3.49 16 17.70 2.36 18.51 3.31 17.80 2.73 21 16.69 2.66 16.90 2.09 16.93 2.78

J. Nutr. Biochem., 1999, vol. 10, November


16:1 (n-7) 12 3.51 1.18 3.59 1.10 3.13 0.80 16 3.69 1.11 4.45* 1.28 4.12 1.28 21 3.19 1.00 3.26 0.83 3.18 0.82
18:0 12 2.61 1.18 2.43 0.97 2.92 1.23 16 2.37 1.17 2.40 1.32 2.16 0.67 21 2.18 1.06 1.95 0.47 2.19 0.93
18:1 (n-9) 12 30.52 3.24 30.55 3.28 32.06 5.87 16 30.63 3.28 29.59 2.78 29.95 3.05 21 28.75 3.55 29.49 2.73 29.15 3.98
18:1 (n-7) 12 2.19 0.38 2.21 0.37 2.41 0.55 16 2.21 0.28 2.34 0.37 2.18 0.47 21 2.02 0.40 1.95 0.30 1.97 0.39
18:2 (n-6) 12 27.12 9.12 29.07 9.19 26.39 10.09 16 29.39 6.78 29.44 7.63 31.22 6.62 21 32.42 7.10 32.96 4.91 31.96 7.20
18:3 (n-3) 12 1.31 0.63 1.67* 0.63 1.86* 0.57 16 1.24 0.50 1.14 0.34 1.24 0.48 21 1.30 0.39 1.30 0.31 1.24 0.33
18:3 (n-6) 12 0.38 0.13 0.46 0.24 0.39 0.32 16 0.41 0.22 0.43 0.16 0.46 0.24 21 0.42 0.13 0.45 0.18 0.46 0.15
18:4 (n-3) 12 0.36 0.09 0.33 0.10 0.30 0.08 16 0.37 0.08 0.35 0.09 0.32 0.08 21 0.32 0.10 0.31 0.08 0.31 0.10
20:1 (n-9) 12 0.23 0.09 0.23 0.11 0.32 0.24 16 0.22 0.08 0.21 0.08 0.20 0.09 21 0.20 0.10 0.18 0.07 0.21 0.08
20:2 (n-6) 12 0.11 0.04 0.11 0.04 0.13 0.05 16 0.10 0.03 0.11 0.06 0.10 0.05 21 0.11 0.05 0.09 0.03 0.11 0.06
20:3 (n-6) 12 0.49 0.23 0.42 0.09 0.37 0.16 16 0.41 0.12 0.46 0.22 0.41 0.08 21 0.47 0.13 0.44 0.07 0.43 0.10
20:4 (n-6) 12 3.26 1.30 3.13 1.09 2.71 1.49 16 3.33 1.01 3.40 0.99 3.28 1.08 21 3.91 1.17 3.99 0.99 3.75 1.13
20:5 (n-3) 12 0.58 0.22 0.67 0.32 0.61 0.39 16 0.64 0.32 0.69 0.55 0.64 0.36 21 1.05 1.25 0.89 0.49 0.88 0.63
22:4 (n-6) 12 0.08 0.22 0.07 0.02 0.08 0.02 12 0.06 0.01 0.06 0.02 0.06 0.02 18 0.06 0.02 0.06 0.02 0.06 0.03
22:5 (n-6) 12 0.08 0.04 0.06 0.02 0.09 0.08 11 0.06 0.03 0.06 0.02 0.06 0.03 15 0.06 0.02 0.06 0.03 0.08 0.05
22:5 (n-3) 12 0.24 0.10 0.25 0.12 0.26 0.13 16 0.20 0.08 0.20 0.09 0.20 0.12 20 0.22 0.11 0.20 0.09 0.22 0.12
22:6 (n-3) 12 0.83 0.43 0.71 0.43 0.70 0.50 16 0.79 0.44 0.79 0.51 0.73 0.47 20 1.11 0.94 0.95 0.50 1.01 0.57

*Significant (P 0.01) compared with point A.


Aat the starting point of the trial. Bafter 1 month of treatment. Cafter 4 months of treatment.
Sea buckthorn oils and atopic dermatitis: Yang et al.

Figure 1 Correlations between symptom improvements and increases in proportions of fatty acids ( 18:3n-3 or 18:2n-6) in plasma lipids in the
seed oil group (a) after 1 month ( SI-AB) and (b) after 4 months of administration ( SI-AC). PL, phospholipids; NL, neutral lipids; A, before treatment;
B, after 1 month of treatment; C, after 4 months of treatment.

pentadecanoic acid (15:0) was decreased by the pulp oil were positively correlated with symptom improvements,
treatment (P 0.01). No significant changes were ob- reflected by a decrease in SCORAD values (correlation
served in the paraffin oil group during follow-up. coefficients and P-values shown in Figure 1). A positive
correlation was also observed between the increase in the
Effects on fatty acid composition of plasma neutral proportion of linoleic acid in plasma phospholipids and AD
lipids symptom improvement, but the correlation was not statisti-
cally significant (Rs 0.36, P 0.28). The overall test of
Twenty-one fatty acids were identified in the plasma neutral the correlation coefficients showed significant positive cor-
lipids and the results are shown in Table 4. At the end of the
relation between symptom improvement and changes in the
administration, the percentage of -linolenic acid increased
proportions of -linolenic and linoleic acids in plasma lipids
in the seed oil group from 1.31 to 1.86%; this was an
caused by seed oil supplementation (P 0.03). In the pulp
increase of over 40%. Pulp oil administration increased the
oil group, no correlations were observed between changes in
percentage of palmitoleic acid and lowered the percentage
SCORAD values and changes in proportions of palmitoleic
of pentadecanoic acid (P 0.01). All the major fatty acid
species such as oleic, linoleic, and palmitic acids remained acid in plasma phospholipids and neutral lipids during the
fairly constant in all participants. Paraffin oil supplementa- follow-up.
tion did not cause changes in the fatty acid composition of
plasma neutral lipids. Effects on the levels of IgE, cholesterol, and
triacylglycerols in plasma
Correlations between symptom improvement and
changes in proportions of fatty acids in plasma The mean levels of plasma IgE, three antigen-specific IgE
antibodies, and plasma lipids of the patients in the three
lipids groups at baseline, after 1 month, and at the end of
In the seed oil group, increases in the proportions of administration are shown in Table 5. Administration of the
-linolenic acid in both phospholipids and neutral lipids three oils did not have significant effects on the levels of

J. Nutr. Biochem., 1999, vol. 10, November 627


Research Communications

total IgE, the three specific IgE antibodies, total cholesterol,

2,566

18.57
SD

6.40

14.4
0.86

0.51
0.43
0.71
8.06
LDL cholesterol, or triacylglycerols in the plasma of the
subjects. A statistically significant, though small increase in
C
Mean

1,997

14.67
13.14

30.48
HDL cholesterol level from the baseline was recognized in

5.08

4.95

1.14
1.49
2.94
the pulp oil group at the end of the treatment. Seed oil and
paraffin oil did not show significant effects on HDL
Table 5 The levels of plasma cholesterol, triacylglycerols, serum total, and specific immunoglobulin E of patients in the three groups at different time points during administration

2,600

21.30
13.28
SD

6.70

0.83

0.43
0.39
0.70
7.61
cholesterol level in plasma.
Placebo
B
Mean

2,125

15.49
14.25

29.91
Discussion
4.59

4.93

1.12
1.45
2.98
In AD patients, deviation in essential fatty acid levels from
2,842
10.16
24.10
18.52
SD

healthy controls have been reported. A higher proportion of


0.74

0.70
0.34
0.63
7.41
linoleic acid and decreased level of arachidonic acid (20:
A

4n-6) have been recognized in plasma lipids.45 47 A com-


Mean

2,485

18.63
16.12

29.60
6.64

4.81

1.14
1.40
2.91
monly accepted hypothesis suggests that AD patients have a
defect in -6 desaturase, which converts linoleic acid and
-linolenic acid to -linolenic acid (18:3n-6) and steridonic
18

10

20

20
20
20
20
n

acid (18:4n-3), respectively. This results in accumulation of


4,708

21.54
11.29
SD

linoleic and -linolenic acids and in decreased levels of


8.21

0.88

0.35
0.35
0.71
7.54

their longer chain desaturated metabolites such as dihomo-


C

-linolenic acid (20:3n-6), arachidonic acid, and eicosapen-


Mean

2,672

11.46
13.99

32.39
1.53*
4.99

4.80

1.01

2.83

taenoic acid (20:5n-3) in plasma phospholipids.47 This


theory is supported by an improvement of AD symptoms,
Sea buckthorn pulp oil

4,340

12.16
16.05
SD

especially itching, after dietary administration of evening


6.85

0.81

0.39
0.28
0.63
6.26

primrose oil rich in -linolenic acid.2 However, Zeven-


B

verger and Houtsmuller48 and Pfeiffer et al.49 demonstrated


Mean

2,410

16.51

30.13

that -6 desaturase is not the only enzyme implicated in the


4.34
7.88

4.43

1.08
1.31
2.63

tissue levels of the PUFA, and they did not find it to be


deficient in AD patients. Thus, more evidence is required to
4,724

20.52
11.69
SD

9.28

0.88

0.30
0.29
0.74
7.29

confirm -6 desaturase deficiency in AD patients.


Clinical studies have shown linoleic acid, the substrate of
A
Mean

2,601

12.49
14.36

30.38

-6 desaturase, to have contradictory effects on AD.6,15,50,51


Aat the starting point of the trial. Bafter 1 month of treatment. Cafter 4 months of treatment.
5.26

4.63

0.96
1.38
2.83

A recently published study showed a significant improve-


ment of AD after fatty acid supplementation in the form of
16

10
16

16
16
16
16
n

8
9

sunflower oil (63% linoleic acid).6 As an essential compo-


nent of the epidermal barrier system, the effect of linoleic
5,400
12.77
16.54
21.11

12.45
SD

0.58

1.17
0.43
0.72

acid was positively correlated with an increased level of


13-hydroxyoctadecadienoic acid (13-HODE) in skin.6 13-
C
Mean

3,027
10.57
13.05
17.35

31.85

HODE, a direct metabolite of linoleic acid, attenuates


4.41

1.48
1.35
2.38

epidermal hyperproliferation6,7,52 and possibly also reduces


Sea buckthorn seed oil

inflammation.7,53 Several clinical trials have also been


1,789
12.07
22.09
20.23

11.56
SD

0.64

1.43
0.39
0.55

carried out to investigate the effects of eicosapentaenoic and


docosahexaenoic acids on AD.5,11 However, -linolenic
B

HDL high density lipoprotein. LDLlow density lipoprotein.


Mean

1,717

14.83
18.56

32.26

acid has attracted less attention in AD treatment.


9.31

4.09

1.52
1.27
2.16

Sea buckthorn seed oil contains 34% linoleic acid and


25% -linolenic acid of its total fatty acids. Supplementa-
1,736
11.19

10.35
SD

*Significant (P 0.05) compared with point A.


17.1
30.8
0.56

1.19
0.38
0.58

tion with the oil resulted in a rise in the proportions of these


two fatty acids in plasma lipids. No increase in the percent-
A

ages of their metabolites -linolenic, dihomo--linolenic,


Mean

1,767

13.91
26.37

32.08
9.41

4.34

1.39
1.36
2.38

arachidonic, or stearidonic acids was observed. However,


the increase in the proportion of -linolenic acid was
12

12
12
12
12
n

accompanied by a slight elevation in the proportion of


9
5
5
7

eicosapentaenoic acid in plasma phospholipids in the seed


mmol/L
mmol/L
mmol/L

oil group, suggesting that -linolenic acid was more effec-


Units

mmol/
IU/L
IU/L
IU/L
IU/L

g/L

tively metabolized in the desaturation-elongation cascade


than was linoleic acid. The results are consistent with the
HDL cholesterol

HDL cholesterol
LDL cholesterol

assumption that -6 desaturation of linoleic acid especially


Triacylglycerols
cholesterol
S. cerevisiae

P. orbiculare

is lowered in AD patients.47
Parameters

C. albicans
Total IgE

Interestingly, in the seed oil group symptom improve-


ments were positively correlated with increase in proportion
Total

of -linolenic acid in plasma lipids caused by seed oil

628 J. Nutr. Biochem., 1999, vol. 10, November


Sea buckthorn oils and atopic dermatitis: Yang et al.

supplementation. The correlation between symptom im- effect shown by pulp oil treatment is probably not related to
provement and increase in proportion of linoleic acid in the fatty acid composition of the oil alone. These results
plasma phospholipids was not significant. This result sug- require confirmation because the number of patients in the
gests positive effects of -linolenic acid on AD. -Linolenic seed oil group was so small. Further studies are in progress
acid is the precursor of eicosapentaenoic acid, which is to discover the effects of the two oil supplementations on
further converted into the 5-series leukotrienes by a 5-li- the fatty acid composition in skin lipids of AD patients.
poxygenase pathway. Eicosapentaenoic acid competitively
inhibits the formation of the 4-series leukotrienes from
arachidonic acid. The 5-series leukotrienes have less potent Acknowledgments
proinflammatory and hyperproliferative effects than the Aromtech Ltd (Tornio, Finland) is acknowledged for pro-
4-series leukotrienes. In our study, supplementation with viding the oil capsules. We thank Ms. Marjukka Sillanpaa
seed oil increased the proportion of eicosapentaenoic acid in and Ms. Terhi Sivonen for participating in part of the fatty
plasma phospholipids. The effects of -linolenic acid may acid analyses.
be due to an elevated synthesis of 5-series leukotrienes and a
decreased synthesis of the 4-series leukotrienes, which resulted
from the increased level of eicosapentaenoic acid in cell References
membranes. However, we did not find a statistically significant
1 Ring, J., Ruzicka, T., and Przybilla, B. (1991). The pathophysiology
correlation between the AD improvement and the increase in of atopic eczema: Synopsis. In Handbook of Atopic Eczema (T.
the proportion of eicosapentaenoic acid in plasma phospholip- Ruzicka, J. Ring, and B. Przybilla, eds.), pp. 330 331, Springer-
ids, possibly due to the low number of patients. Future studies Verlag, Berlin, Germany
on the effects of seed oil supplementation on the levels of 2 Morse, P.F., Horrobin, D.F., Manku, M.S., Stewart, J.C.M., Allen,
-linolenic acid, linoleic acid, and their metabolites in skin R., Littlewood, S., Wright, S., Burton, J., Gould, D.J., Holt, P.J.,
Jansen, C.T., Mattila, L., Meigel, W., Dettke, T.H., Wexler, D.,
lipids should provide valuable information of the function and Guenther, L., Bordoni, A., and Patrizi, A. (1989). Meta-analysis of
metabolism of essential fatty acids in AD patients. placebo-controlled studies of the efficacy of Epogam in the treatment
Significant improvement also occurred in the paraffin oil of atopic eczema. Relationship between plasma essential fatty acid
group without any changes in plasma fatty acid levels, changes and clinical response. Brit. J. Dermatol. 121, 7590
3 Burton, J.L. (1990). Essential fatty acids in atopic eczema: Clinical
suggesting the improving effect of the traditional treat- studies. In Omega-6 Essential Fatty Acids: Pathophysiology and
ments. This may have been due to the better compliance Roles in Clinical Medicine (D.F. Horrobin, ed.), pp. 6773, Alan R.
provoked by the regular control visits during the trial. This Liss, Inc., New York, NY, USA
kind of improvement, the placebo effect, was also high- 4 Bahmer, F.A. and Schafer, J. (1992). Die Behandlung der atopischen
Dermatitis mit Borretschsamen-O l (Glandol) - eine zeitreihenana-
lighted by Morse et al.2 in the meta-analysis of 9 placebo-
lytische Studie. Kinderarztl. Praxis 60, 199 202
controlled studies on the efficacy of evening primrose oil in 5 Syland, E., Funk, J., Rajka, G., Sandberg, M., Thune, P., Rustad, L.,
the treatment of AD. At the starting point of the trial, the Helland, S., Middelfart, K., Odu, S., Falk, E.S., Solvoll, K., Bjrne-
general symptom severity in the seed oil group happened to boe, G.E.A., and Drevon, C.A. (1994). Dietary supplementation with
be lower than in the pulp oil and placebo groups. Together very long-chain n-3 fatty acids in patients with atopic dermatitis. A
double-blind, multicenter study. Brit. J. Dermatol. 130, 757764
with the smaller number of patients, this may explain the 6 Gimenez-Arnau, A., Barranco, C., Alberola, M., Wale, C., Serrano,
less significant symptom improvement in the seed oil group S., Buchanan, M.R., and Camarasa, J.G. (1998). Effects of linoleic
compared with the other two groups. acid supplements on atopic dermatitis. In Recent Advances in
The high content of palmitoleic acid in the pulp oil Prostaglandin, Thromboxane, and Leukotriene Research (Sinzinger
resulted in an increase in the proportion of the fatty acid in et al. eds.), pp. 285289, Plenum Press, New York
7 Ziboh, V.A. (1996). The significance of polyunsaturated fatty acids
plasma lipids. The increase neither correlated with the in cutaneous biology. Lipids 31, S249 253
symptom improvement nor led to clear changes in the 8 The British Nutrition Foundation (1992). Function of unsaturated
plasma levels of total cholesterol, LDL cholesterol, or fatty acids. In Unsaturated Fatty Acids: Nutrition and Physiological
triacylglycerols. The significant improving effect of the Significance (The Report of The British Nutrition Foundations Task
Force), pp. 48 62, Chapman & Hall, London, UK
pulp oil also may be due to components other than its fatty 9 Horrobin, D.F. and Manku, M.S. (1990). Clinical biochemistry of
acids. According to our analysis (unpublished results), pulp essential fatty acids. In Omega-6 Essential Fatty Acids: Pathophys-
oil contains a higher amount of plant sterols (2%) than seed iology and Roles in Clinical Medicine (D.F. Horrobin, ed.), pp.
oil (0.8%). In the pulp oil plant sterol fraction, 68% consists 2150, Alan R. Liss, Inc., New York, NY, USA
of sitosterol and 5% of sitostanol. Both of these are reported 10 Syland, E., Nenseter, M.S., Braathen, L., and Drevon, C.A. (1993).
Very long chain n-3 and n-6 polyunsaturated fatty acids inhibit
to have an anti-inflammatory effect.54 56 Zak et al.57 re- proliferation of human T-lymphocytes in vitro. Eur. J. Clin. Invest.
ported that dietary supplementation with sitosterol de- 23, 112121
creased LDL and slightly increased HDL levels in hyper- 11 Bjrneboe, A., Syland, E., Bjrneboe, G.E.A., Rajka, G., and
cholesterolemic patients. The slight increase in HDL level Drevon, C.A. (1987). Effect of dietary supplementation with eico-
sapentaenoic acid in the treatment of atopic dermatitis. Brit. J.
discovered after the pulp oil treatment in our study is in Dermatol. 117, 463 469
agreement with their observations. The high content of 12 Biagi, P.L., Bordoni, A., Hrelia, S., Celadon, M., Ricci, G.P.,
carotenoides (1 mg/g oil), especially of -carotene (0.4 Cannella, V., Patrizi, A., Specchia, F., and Masi, M. (1994). The
mg/g oil), which is an antioxidant and precursor of vitamin effect of -linolenic acid on clinical status, red cell fatty acid
A, in the pulp oil may also have contributed to the composition and membrane microviscosity in infants with atopic
dermatitis. Drug. Exp. Clin. Res. 20, 77 84
significant AD improvement in the pulp oil group. 13 Andreassi, M., Forleo, P., Di Lorio, A., Masci, S., Abate, G., and
In conclusion, -linolenic acid in sea buckthorn seed oil Amerio, P. (1997). Efficacy of -linolenic acid in the treatment of
seemed to have a beneficial effect on AD. The improving patients with atopic dermatitis. J. Int. Med. Res. 25, 266 274

J. Nutr. Biochem., 1999, vol. 10, November 629


Research Communications
14 Berth-Jones, J. and Graham-Brown, R.A.C. (1993). Placebo-con- microbes in supercritical CO2 extraction of sea buckthorn (Hippo-
trolled trial of essential fatty acid supplementation in atopic derma- phae rhamnoides) oils. Z. Lebensm. Unters. For. 204, 202205
titis. Lancet 341, 15571560 37 European Task Force on Atopic Dermatitis. (1993). Severity scoring
15 Bamford, J.T., Gibson, R.W., and Renier, C.M. (1985). Atopic of atopic dermatitis: the SCORAD index. (Consensus Report of the
eczema unresponsive to evening primrose oil (linoleic and gamma- European Task Force on Atopic Dermatitis). Dermatology 186,
linolenic acids). J. Am. Acad. Dermatol. 13, 959 965 2231
16 Chardon, P., Crastes de Paulet, P., Lemat, A.M., Crastes de Paulet, 38 Folch, J., Lees, M., and Sloane-Stanley, G.H. (1957). A simple
A., Kienlen, J., and du Cailar, J. (1985). Effet dune alimentation method for isolation and purification of total lipids in normal man.
enterale exclusive par Enteronutril sur les lipides du serum. Ann. J. Biol. Chem. 226, 497509
Fr. Anesth. 4, 333338 39 Ways, P. and Hanahan, D.J. (1964). Characterization and quantifi-
17 Berghaus, G., Kurz, A., and Dotzauer, G. (1975). Vergleich des cation of red cell lipids in normal man. J. Lipid Res. 5, 318 328
Fettsauremusters zwischen menschlichem Fett- und Lebergewebe 40 Christie, W.W. (1982). A simple procedure for rapid transmethyla-
bei Nichtalkoholikern und Alkoholikern. Dtsch. med. Wschr. 100, tion of glycerolipids and cholesteryl esters. J. Lipid Res. 23,
12331238 10721075
18 Gomez-Tubo, A., Pita, M.L., Tavares, E., Murillo, M.L., Delgado, 41 Izzo, C., Grillo, F., and Murador, E. (1981). Improved method for
M.J., and Carreras, O. (1995). Changes in the fatty acid profile of determination of high-density-lipoprotein cholesterol. I. Isolation of
plasma and adipose tissue in rats after long-term ethanol feeding. high-density-lipoproteins by use of polyethylene glycol 6000. Clin.
Alcohol. Clin. Exp. Res. 19, 747752 Chem. 27, 371374
19 Colquhoun, D.M., Humphries, J.A., Moores, D., and Somerset, S.M. 42 Friedewald, W.T., Levy, R.I., and Fredrickson, D.S. (1972). Estima-
(1996). Effects of a macadamia nut enriched diet on serum lipids and tion of the concentration of low-density-lipoprotein cholesterol in
lipoproteins compared to a low fat diet. Food. Aust. 48, 216 222 plasma, without use of the preparative ultracentrifuge. Clin. Chem.
20 Zhou, Y. and Jiang, J. (1989). Medical and health-care functions and 18, 499 502
applications of sea buckthorn. Hippophae (Chinese) 2(2), 35 42 43 Pharmacia & Upjohn Diagnostics AB (Uppsala, Sweden) (1997).
21 Quirin, K.W. and Gerard, D. (1993). Sanddornlipideinteressante Wirk- Pharmacia IgE RIA: Direction for Use.
stoffe fur die Kosmetik. Parfumerie und Kosmetik 10/1993, 618 625 44 Pharmacia & Upjohn Diagnostics AB (Uppsala, Sweden) (1997).
22 Berezhnaya, G.A., Ozerinina, O.V., Yeliseev, I.P., Tsydendambaev, Pharmacia CAP SystemTM RAST RIA: Direction for Use.
V.D., and Vereshchagin, A.G. (1993). Developmental changes in the 45 Manku, M.S., Horrobin, D.F., Morse, N.L., Wright, S., and Burton,
absolute content and fatty acid composition of acyl lipids of sea J.L. (1984). Essential fatty acids in the plasma phospholipids of
buckthorn fruits. Plant Physiol. Biochem. 31(3), 323332 patients with atopic eczema. Brit. J. Dermatol. 110, 643 648
23 Johansson, A., Laakso, P., and Kallio, H. (1997). Characterization of 46 Strannegrd, I.L., Svennerholm, L., and Strannegrd, O . (1987).
seed oils of wild, edible Finnish berries. Z. Lebensm. Unters. For. Essential fatty acids in serum lecithin of children with atopic
A204, 300 307 dermatitis and in umbilical cord serum of infants with high or low
24 Chen, Y., Jiang, Z., Qin, W., Ni, M., Li, X., and He, Y. (1990). IgE levels. Int. Archs. Allergy appl. Immun. 82, 422 423
Chemical composition and characteristics of sea buckthorn fruit and 47 Wright, S. (1990). Essential fatty acids and atopic eczema: Biochem-
its oil. Chemistry and Industry of Forest Products (Chinese) 10(3), ical and immunological studies. In Omega-6 Essential Fatty Acids:
163175 Pathophysiology and Roles in Clinical Medicine (D.F. Horrobin,
25 Xin, Y., Zhou, P., Chang, J., Zhao, X., Wang, C., and Ren, J. (1993). ed.), pp. 55 65, Alan R. Liss, Inc., New York, NY, USA
Characteristics of sea buckthorn oils and research on the oil 48 Zevenbergen, J.L. and Houtsmuller, U.M.T. (1989). Effects of
processing technology. China Oil and Fat (Chinese) 2/1993, 8 11 dietary fats on linoleic acid metabolism. A radiolabel study in rats.
26 Mironov, V.A., Vasilev, G.S., Matrosov, V.S., Muzychenko, L.D., Biochim. Biophys. Acta 1002, 312323
Usha, B.V., Kasyanenko, I.I., and Feldshtein, M.A. (1980). Sea 49 Pfeiffer, A., Demmelmair, H., Rueff, F., Przybilla, B., Reinhardt, D.,
buckthorn oil obtained by the extraction method and its biological and Koletzko, B. (1996). Normal in vivo activity of -6 desaturase
activity. Khim. Farm Zh. (Russian) 14(8), 74 80 in patients with atopic eczema. International Conference on Highly
27 Mironov, V.A., Guseva-Donskaya, T.N., Dubrovina, Y., Osipova, Unsaturated Fatty Acids in Nutrition and Disease Prevention,
G.A., Shabanova, E.A., Nikulin, A.A., Amirov, N.S., and Trubitsina, Barcelona, Spain, November 4 6
I.G. (1991). Composition and biological activity of lipid extracts 50 Hansen, A.E. (1933). Serum lipid changes and therapeutic effects of
from Armenian sea buckthorn. In Nov. Biol. Khimii I Farmakol. various oils in infantile eczema. Proceedings of the Society for
Oblepikhi. (Russian) pp. 114 121, SO AN SSSR, Novosibirsk, Experimental Biology and Medicine 31, 160 161
Russia 51 Cornbleet, T. (1935). The use of maize oil (unsaturated fatty acids)
28 Li, Y. and Xu, M. (1993). Preliminary report on the anti-bacterial in the treatment of eczema. Arch. Dermatol. Syphilol. (Chicago) 31,
effect of sea buckthorn oil. Hippophae (Chinese) 6(2), 28 29 224 234
29 Zhao, Y. (1994). Preliminary report on the effects of sea buckthorn 52 Miller, C.C. and Ziboh, V.A. (1990). Induction of epidermal hyper-
oil on thirty-two cases of burn or scald. Hippophae (Chinese) 7(3), proliferation by topical n-3 polyunsaturated fatty acids on guinea pig
36 37 skin linked to decreased levels of 13-hydroxyoctadecadienoic acid
30 Zheng, H., Chen, X., Yang, Q., and He, F. (1990). The effect of (13-HODE). J. Invest. Dermatol. 94, 353358
Hippophae rhamnoides seed oil on immune function of mice. 53 Buckley, T.L., Van de Velde, M.J., Henricks, P.A., Engels, F., and
Academic Journal of Lanzhou University (Chinese) 26(2), 9598 Nijkamp, F.P. (1992). 13-Hydroxyoctadecadienoic acid attenuates
31 Hasigerile and Wu, Y. (1993). Effect of Hippophae rhamnoides oil oedema formation induced by leukotriene B4 in vivo in rabbit skin.
on peritoneal macrophage in mice. Academic Journal of Inner Eur. J. Pharmacol. 218(23), 369 372
Mongolia Medical University (Chinese) 15(1), 30 32 54 Yasukawa, K., Akihisa, T., Kimura, Y., Tamura, T., and Takido, M.
32 Xu, W., Che, X., Huang F., and Che, X. (1997). Effect of sea (1998). Inhibitory effect of cycloartenol ferulate, a component of rice
buckthorn seed oil on immune function of mice. Hippophae (Chi- bran, on tumor promotion in two-stage carcinogenesis in mouse skin.
nese) 10(3), 29 32 Biol. Pharm. Bull. 21(10), 10721076
33 Ji, Y. and Gao, Y. (1991). Effect of feeding of sea buckthorn oil and 55 Yasukawa, K., Takido, M., Matsumoto, T., Takeuchi, M., and
sea buckthorn oil supplemented with sodium selenite in vivo on Nakagawa, S. (1991). Sterol and triterpene derivatives from plants
structural stability of erythrocyte membrane in rats. Journal of inhibit the effects of a tumor promoter, and sitosterol and betulinic
Biochemistry (Chinese) 7, 441 446 acid inhibit tumor formation in mouse skin two-stage carcinogenesis.
34 Song, Z. and Gao, Y. (1995). Effect of sea buckthorn oil and vitamin Oncology 48(1), 7276
E on lipid peroxidation of rat after cold exposure. Acta Nutrimenta 56 Jiang, Z. and Li, G. (1987). Study on the active component of sea
Sinica (Chinese) 17(1), 2731 buckthorn oil for treating the gastric ulcer. Academic Journal of the
35 Jiang, Y., Zhou, Y., Bi, C., Li, J., Yang, J., Yu, Z., Hu, Z., and Zhao, Second Military Medical University of China (Chinese) 8(2), 119
S. (1993). Study on the clinical effect of sea buckthorn oil on 57 Zak, A., Zeman, M., Vitkova, D., Hrabak, P., and Tvrzicka, E.
hyperlipaemia. Hippophae (Chinese) 6(3), 2324 (1990). Beta-sitosterol in the treatment of hypercholesterolemia.
36 Manninen, P., Haivala, E., and Kallio, H. (1996). Distribution of Cas. Lek. Cesk. 129, 1320 1323

630 J. Nutr. Biochem., 1999, vol. 10, November